Characterization and Early Assays Development in FOXG1 Deficient Neurons

项目摘要 FOXG 1综合征是一种罕见的神经系统疾病，通常会导致严重的智力残疾， 小头畸形和神经系统症状，包括张力减退、癫痫发作和刻板 动作FOXG 1综合征是由叉头基因杂合突变或缺失引起的 盒1蛋白（FOXG 1），一种控制关键皮质发育表达的转录因子 proteins. FOXG 1对端脑神经前体细胞（NPC）具有特异性重要作用 增殖和分化为GABA能中间神经元。我们优先考虑FOXG 1综合征 基于我们开发的评估单基因神经发育障碍的框架， 基于仿制药、临床前验证、临床和伦理的治疗开发 注意事项。我们还从FOXG 1中创建了三个多能干细胞（PSC）系 综合征患者，并使用CRISPR/Cas9基因编辑来创建同基因对照系。我们 假设一种药物样的小分子，增加FOXG 1表达的患者， 产后早期发育将改善这些人的症状。拟议 研究中，我们将使用FOXG 1 PSC系通过以下方式创建内源位点基因报告系： 使用CRISPR/Cas9在FOXG 1基因的C末端插入HaloTag报告蛋白。 将开发一种方法，从而使这些修饰的FOXG 1 PSC分化为 皮质神经祖细胞（NPC）并用HaloTag配体系留染料处理以测量 准确测量FOXG 1表达。该项目的主要目标是展示 这种FOXG 1报告系统适用于筛选小分子， 增加FOXG 1表达。

PROJECT SUMMARY FOXG1 syndrome is a rare neurological disorder that often causes severe intellectual disability, microcephaly and neurological symptoms including hypotonia, seizures, and stereotypic movements. FOXG1 syndrome is caused by heterozygous mutation or deletion of the forkhead box 1 protein (FOXG1), a transcription factor that controls expression of key cortical development proteins. FOXG1 is specifically important for telencephalon neural progenitor cell (NPC) proliferation and differentiation to GABAergic interneurons. We have prioritized FOXG1 syndrome based on a framework we developed to evaluate monogenetic neurodevelopmental disorders for therapeutic development based on generic, preclinical validation, clinical and ethical considerations. We have also created three pluripotent stem cell (PSC) lines from FOXG1 syndrome patients and used CRISPR/Cas9 gene editing to create isogenic control lines. We hypothesize that a drug-like small molecule that increases FOXG1 expression in patients during early postnatal development would improve symptoms in these individuals. In the proposed research, we will use FOXG1 PSC lines to create an endogenous locus gene reporter line by using CRISPR/Cas9 to insert a HaloTag reporter protein at the C-terminus of the FOXG1 gene. A method will be developed whereby these modified FOXG1 PSCs with be differentiated to cortical neural progenitor cells (NPCs) and treated with HaloTag ligand tethered dyes to measure accurately measure FOXG1 expression. The primary goal of this project will be to demonstrate that this FOXG1 reporter system is suitable for screening small molecules for their ability to increase FOXG1 expression.